GR Luciferase Reporter MDA-MB-453 Stable Cell Line

GR Responsive Luciferase Reporter MDA-MB-453 Stable Cell Line is derived from human breast cancer, and stably express firefly luciferase reporter gene under the control of the GR response element. This cell line is an ideal cellular model for monitoring the activation of Glucocorticoid Receptor Signaling Pathway triggered by stimuli treatment, enforced gene expression and gene knockdown.

Principle

The Glucocorticoid receptor (GR) is a major player in development, metabolism and immune response. When activated by stimuli, GR translocates from the cytoplasm into the nucleus, and then binds a DNA recognition site to regulate gene expression. In addition, activated GR can transrepress other transcription factors that have been misregulated in cancer and other diseases.

The cell line was established by transfection of a GR firefly luciferase reporter vector along with G418 expression vector followed by G418 selection. The G418 resistant clones were subsequently screened for DEX-induced luciferase activity. The clone with the highest fold induction (38 fold) was selected and expanded to produce this stable cell line.

Principle behind TF luciferase reporter. TF luciferase reporter stable cell line utilizes artificial promoter constructs to drive luciferase expression. The promoter region can consists of multiple repeats of a cis-element TF binding site, a DNA fragment from the promoter region of a known TF downstream gene, or a DNA fragment containing putative/known TF binding sites. There are several ways that a TF can be activated, such as through extracellular stimuli or through intracellular signaling pathways. Once activated, the TF translocates to the nucleus and often interacts with relevant co-factors to drive gene expression. Once luciferase is expressed, it can generate light in an enzymatic assay and the amount of light measured is positively correlated with the level of TF activation.

Data

Analysis of GR pathway reporter MDA-MB-453 cell line in response to stimuli. The cells were seeded on a 96-well plate for overnight with DMEM including 10% FBS. The cells then were treated with or without 1uM Dexamethasone (DEX) respectively in DMEM and 0.1% FBS for 16-24 hours.